Improved Diagnostics and Individualized Therapeutics using Optical Microring Reso

描述（由申请人提供）：拟议研究的目的是使用光学微共振生物分析平台来分析模型细胞系和现实生活中多形性胶质母细胞瘤（GBM）肿瘤活检样本中存在的磷酸化蛋白。这种多参数、无标记技术的固有优势，具有增加的灵敏度，寻求通过减少样本量要求来提高当前的诊断能力，同时在个体基础上评估导致GBM发展和进展中所涉及的信号转导途径（STPs）失调的翻译后修饰。磷酸化特异性抗体和光学生物分析平台旨在通过用侵入性较低的皮肤针活检替代石蜡包埋肿瘤活检样本的标准免疫组织化学分析来降低样本获取程序的严重性。该平台还将通过确定各种STP的失调并为患者引入正确的抑制剂来增加GBM治疗程序的特异性。 本研究提出了使用光学生物分析平台，确定在六个模型细胞系中的相对蛋白质表达，这些细胞系被转染为具有PTEN、EGFR和EGFRvIII（单独和组合）的过表达。该研究计划随后扩展到监测涉及来自相同六个细胞系的蛋白质如PI 3 K、Akt、mTOR和MAPK/ERK的其他下游信号转导级联。还将在这些相同的细胞系上研究单个蛋白激酶以及单个和组合疗法的影响。最后，本研究预期监测磷蛋白的能力，有效地预测信号STP失调，负责从石蜡包埋的肿瘤活检组织中获得的患者的GBM的发展。 公共卫生相关性：拟议的工作将通过提高诊断能力和降低获得肿瘤活检的侵入性来有益于公共卫生，并有望提高治疗多形性胶质母细胞瘤的个体化疗法的疗效。本研究旨在弥合GBM发生和发展的信号转导通路失调的分子起源之间的差距。光学微型共振器生物分析平台的集成沿着磷酸化蛋白质组学的最新进展，不仅可以确定“这是癌症吗？”更早、侵入性更小，但也将提供一种方法来回答“我如何最有效地治疗这种疾病？”'

DESCRIPTION (provided by applicant): The objective of the proposed research is to implement the use of an optical microring resonator bioanalysis platform to profile the phosphorylated proteins present in model cell lines and real-life glioblastoma multiforme (GBM) tumor biopsy samples. The inherent advantages of this multiparameter, label-free technology, with increased sensitivity, seeks to advance current diagnostic capabilities by reducing sample size requirements while simultaneously, evaluating on an individual basis, post-translational modifications that lead to the deregulation of signal transduction pathways (STPs) involved in the development and progression of GBM. Incorporating phospho-specific antibodies and the optical bioanalysis platform aims to reduce the severity of the procedure for sample obtainment by replacing standard immunohistochemical analysis of paraffin-embedded tumor biopsy samples with less invasive skinny-needle biopsies. This platform will also serve to increase the specificity of GBM treatment procedures by determining the deregulation of various STPs and incorporating the right inhibitor(s) for the patient. This research proposes to determine relative protein expression in six model cell lines transfected to have over-expression of PTEN, EGFR and EGFRvIII (separately and in combination), using the optical bioanalysis platform. This research plan is then expanded to monitor other downstream signal transduction cascades involving proteins such as PI3K, Akt, mTOR and MAPK/ERK from the same six cell lines. The affect of individual protein kinases, as well as individual and combinatorial therapies will also be studied on these same cell lines. Finally, this research anticipates the ability to monitor the phosphoproteins, effectively predicting the signal STP deregulation, responsible for the development of GBM in patients from which paraffin-embedded tumor biopsy tissues are obtained. PUBLIC HEALTH RELEVANCE: The proposed work will benefit the public health by improving diagnostic capabilities and decreasing the invasiveness of obtaining tumor biopsies, with the promise to increase the efficacy of individualized therapeutics for the treatment of glioblastoma multiforme. This research intends to bridge the gap between the molecular origins for the deregulation of signal transduction pathways resulting in the development and progression of GBM. The integration of an optical microring resonator bioanalysis platform along with the recent advancements in phosphoproteomics will not only determine 'is this cancer?' earlier and less invasively, but will also provide a way to answer the question of 'how do I most effectively treat this disease?'